hTERT Human Prostate Fibroblasts

$6,000.00

Puromycin-resistant hTERT-Cells are generated by infecting Human Prostate Cancer-Associated Fibroblasts (HPCAF) with lentiviruses expressing hTERT, followed by antibiotic selection. These primary Human Prostate CAFs are isolated directly from human prostate tumor tissue and cultured in T75 flasks pre-coated with AlphaBioCoat solution for 30 minutes before being maintained in CAF Growth Medium for 3–7 days. After initial expansion, the cells are cryopreserved at passage 1, with each vial containing a minimum of 1,000,000 cells per milliliter for shipment.

Puromycin-resistant hTERT-Cells are generated by infecting Human Prostate Cancer-Associated Fibroblasts (HPCAF) with lentiviruses expressing hTERT, followed by antibiotic selection. These primary Human Prostate CAFs are isolated directly from human prostate tumor tissue and cultured in T75 flasks pre-coated with AlphaBioCoat solution for 30 minutes before being maintained in CAF Growth Medium for 3–7 days. After initial expansion, the cells are cryopreserved at passage 1, with each vial containing a minimum of 1,000,000 cells per milliliter for shipment.

Product Testing & Quality Control: Human Prostate Cancer-Associated Fibroblasts (CAFs) undergo rigorous characterization, confirming negative expression of endothelial (von Willebrand Factor/Factor VIII), epithelial (cytokeratin 18), and smooth muscle (α-SMA) markers while testing negative for microbial contaminants including bacteria, yeast, fungi, and mycoplasma. These cells positively express established CAF biomarkers (FAP, PDGFR, Vimentin, PDPN, and CD70) and maintain stable growth for 3-5 passages at recommended 1:2-1:3 split ratios.

Research Applications: Suitable for diverse experimental protocols including cell-cell interaction studies, adhesion assays, molecular biology techniques (PCR, Western blot, immunoprecipitation), fluorescence-based analyses (immunofluorescence, flow cytometry), and generation of specialized cellular models.

Cell Handling & Culture Protocols:

  • Thawing Procedure: For optimal recovery, rapidly thaw frozen vials (2 min in 37°C water bath), sterilize with 70% ethanol, and process under sterile conditions. Immediately remove cryoprotectant via centrifugation (125 × g, 5-10 min) and resuspend in fresh medium. Pre-coat culture vessels with AlphaBioCoat (6-8 mL, 15 min) followed by PBS rinse before plating. Pre-equilibrate medium (15 min at 37°C, 5% CO₂) to stabilize pH (7.0-7.6) prior to cell seeding.

  • Subculture Protocol: Detach confluent cells using PBS rinse and enzymatic treatment (2-3 mL detachment solution, 5-15 min at 37°C if needed). Re-coat flasks with AlphaBioCoat before reseeding at 1:2-1:3 ratios in fresh medium (renewed every 3-4 days). Avoid mechanical agitation to prevent clumping.

  • Cryopreservation: Use complete growth medium with 5% DMSO (Catalog #CGM5) for cell freezing.

Storage & Shipping: Maintain frozen vials in liquid nitrogen vapor phase; avoid -70°C storage to preserve viability. For extended storage post-thawing, return cells to cryogenic conditions promptly.